                         SEQUENCE LISTING

<110>  Zymergen Inc.
 
<120>  METHOD FOR COUNTERSELECTION IN MICROORGANISMS

<130>  ZYMR-048/01WO 327574-2310

<150>  US 62/904,285
<151>  2019-09-23

<160>  3     

<170>  PatentIn version 3.5

<210>  1
<211>  1035
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  codon optimized PheS(**CO) counterselection marker

<400>  1
atggaagaga agcttaaaca gctcgaactg gaagctgcag aaaaagtaga agcggctggt       60

tcattaaaag aagttaacga tattcgggtg caatatctcg gaaaaaaagg gccgattaca      120

gaagttctcc gcggcatggg caaactgtcc gccgaagagc gcccgaaaat gggagccctc      180

gccaatgaag tgagagagcg cattgcagca gccataacgg cgaaaaatga acagcttgaa      240

caagaagaga tgaataaaaa actcagcagt cagacgattg atgtcacatt gcctggcagt      300

ccggtaaaca tcggcggacg ccatccgctg acggtcgtta ttgaagaaat tgaagattta      360

tttatcggta tgggatatac cgtagaagaa ggccctgaag tggaaacgga ttattataac      420

ttcgaagcgc tgaatctgcc gaaggaacat ccggcccgcg acatgcagga cagcttttac      480

attacagaag aaatgctgat gagaacgcag acgtcaccgg tgcagacacg cacgatggaa      540

aaacataaag gaaaaggccc cgtcaaaatc atttgtcccg gtaaagtgta tcgccgtgac      600

aacgatgacg cgacacattc ccatcagttc atgcagatcg agggtctcgt cgtggaccgc      660

aaaatcagca tgagcgacct gaaaggcacg cttgaattgg tcgcgaaaaa aatgttcggc      720

caagaccgtg aaatcaggct ccgtccgagc ttcttcccat tctccgagcc ttctgtcgaa      780

gtggatgtga cctgctttaa gtgcggcgga caaggctgct ccgtctgcaa aaaaacaggc      840

tggatcgaaa ttctcggcgc cggcatggtt cacccgaacg tgctgaaaat ggccgggttc      900

aatcctgaag agtaccaggg ttttggcttc ggaatgggtg ttgaacgtat cgcgatgctg      960

aaatacggca ttgaagatat ccgccatttc tataccaatg atgtcagatt catttcgcag     1020

tttaaacagg cgtaa                                                      1035


<210>  2
<211>  1035
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  codon optimized PheS(**) counterselection marker

<400>  2
atggaagaga aactcaaaca gcttgagctg gaagcagccg agaaagtaga ggcggcgggt       60

agcctcaaag aggtgaacga cattcgggta caatacctcg gtaagaaagg cccaataaca      120

gaagttttga gaggaatggg gaaactcagt gctgaggaga gaccgaaaat gggggcgctc      180

gccaacgaag ttcgggaaag aatcgcagca gctataaccg cgaaaaatga acaacttgaa      240

caagaagaaa tgaataagaa attgtcttct caaacgatcg acgttactct cccaggatca      300

ccggtcaata taggcgggag acatccgctc acggtcgtga tagaggaaat cgaagacctc      360

tttatcggga tgggttatac ggtggaagaa ggacctgagg tcgaaactga ttattacaat      420

ttcgaggctt taaatctgcc aaaagaacac cctgcgcggg atatgcagga ttctttttac      480

atcacggagg agatgctcat gagaactcag actagtcctg tgcaaacgcg gacgatggaa      540

aagcacaaag gaaaggggcc tgtcaagatc atctgtccgg ggaaggtcta ccggcgggat      600

aacgacgacg cgactcatag tcatcaattc atgcaaattg agggcttggt agtggaccgt      660

aaaatttcca tgtctgattt gaaggggact cttgaattag ttgctaagaa aatgtttggt      720

caagacagag agatacgcct ccgcccatca ttttttccgt ttagtgagcc aagcgtagag      780

gtagatgtta cgtgcttcaa atgtggcggc caggggtgca gcgtgtgtaa gaagaccggc      840

tggattgaga tattaggggc cggaatggta catccaaatg tgttgaagat ggctggtttc      900

aacccagaag agtatcaagg cttcggattc gggatggggg tagagcgtat tgcaatgctt      960

aagtatggta tcgaagacat acgccatttc tatacaaatg acgttcgctt catttctcag     1020

ttcaaacagg cgtaa                                                      1035


<210>  3
<211>  344
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alpha-subunit of phenylalanyl-tRNA ligase (PheS) with A309G/T255S mutations 

<400>  3

Met Glu Glu Lys Leu Lys Gln Leu Glu Leu Glu Ala Ala Glu Lys Val 
1               5                   10                  15      


Glu Ala Ala Gly Ser Leu Lys Glu Val Asn Asp Ile Arg Val Gln Tyr 
            20                  25                  30          


Leu Gly Lys Lys Gly Pro Ile Thr Glu Val Leu Arg Gly Met Gly Lys 
        35                  40                  45              


Leu Ser Ala Glu Glu Arg Pro Lys Met Gly Ala Leu Ala Asn Glu Val 
    50                  55                  60                  


Arg Glu Arg Ile Ala Ala Ala Ile Thr Ala Lys Asn Glu Gln Leu Glu 
65                  70                  75                  80  


Gln Glu Glu Met Asn Lys Lys Leu Ser Ser Gln Thr Ile Asp Val Thr 
                85                  90                  95      


Leu Pro Gly Ser Pro Val Asn Ile Gly Gly Arg His Pro Leu Thr Val 
            100                 105                 110         


Val Ile Glu Glu Ile Glu Asp Leu Phe Ile Gly Met Gly Tyr Thr Val 
        115                 120                 125             


Glu Glu Gly Pro Glu Val Glu Thr Asp Tyr Tyr Asn Phe Glu Ala Leu 
    130                 135                 140                 


Asn Leu Pro Lys Glu His Pro Ala Arg Asp Met Gln Asp Ser Phe Tyr 
145                 150                 155                 160 


Ile Thr Glu Glu Met Leu Met Arg Thr Gln Thr Ser Pro Val Gln Thr 
                165                 170                 175     


Arg Thr Met Glu Lys His Lys Gly Lys Gly Pro Val Lys Ile Ile Cys 
            180                 185                 190         


Pro Gly Lys Val Tyr Arg Arg Asp Asn Asp Asp Ala Thr His Ser His 
        195                 200                 205             


Gln Phe Met Gln Ile Glu Gly Leu Val Val Asp Arg Lys Ile Ser Met 
    210                 215                 220                 


Ser Asp Leu Lys Gly Thr Leu Glu Leu Val Ala Lys Lys Met Phe Gly 
225                 230                 235                 240 


Gln Asp Arg Glu Ile Arg Leu Arg Pro Ser Phe Phe Pro Phe Ser Glu 
                245                 250                 255     


Pro Ser Val Glu Val Asp Val Thr Cys Phe Lys Cys Gly Gly Gln Gly 
            260                 265                 270         


Cys Ser Val Cys Lys Lys Thr Gly Trp Ile Glu Ile Leu Gly Ala Gly 
        275                 280                 285             


Met Val His Pro Asn Val Leu Lys Met Ala Gly Phe Asn Pro Glu Glu 
    290                 295                 300                 


Tyr Gln Gly Phe Gly Phe Gly Met Gly Val Glu Arg Ile Ala Met Leu 
305                 310                 315                 320 


Lys Tyr Gly Ile Glu Asp Ile Arg His Phe Tyr Thr Asn Asp Val Arg 
                325                 330                 335     


Phe Ile Ser Gln Phe Lys Gln Ala 
            340                 


